基于单细胞RNA测序的人羊水间充质干细胞异质性分析  

Heterogeneity analysis of human amniotic fluid mesenchymal stem cells based on single cell RNA sequencing

作　　者：郑玉婷 朱晓帆[1] 杨宇霞[2] 郭瑞霞[3] 孔祥东[1] ZHENG Yuting;ZHU Xiaofan;YANG Yuxia;GUO Ruixia;KONG Xiangdong(Center of Genetic and Prenatal Diagnosis,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Department of Emergency,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Department of Gynecology and Obstetrics,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)

机构地区：[1]郑州大学第一附属医院遗传与产前诊断中心,郑州450052 [2]郑州大学第一附属医院急诊内科,郑州450052 [3]郑州大学第一附属医院妇产科,郑州450052

出　　处：《郑州大学学报（医学版）》2022年第4期455-461,共7页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省医学科技攻关计划项目(LHGJ20190075)。

摘　　要：目的:利用单细胞RNA测序技术(scRNA-seq)确定人羊水间充质干细胞(AF-MSC)的细胞亚群及各个亚群之间的异质性。方法:采集孕中期正常孕妇羊水样本,分离培养获得AF-MSC,使用10x scRNA-seq平台进行检测。对测序结果进行过滤、比较、量化、筛选等分析,根据基因表达情况进行降维聚类分群并判断细胞周期,对亚群进行差异基因分析并获得标记基因,预测不同亚组的分化能力,分析表面标志物的表达水平。结果:共检测到6717个细胞,根据基因表达情况将细胞聚类分为7个亚群。亚群4为非MSC,余亚群均为MSC。细胞周期影响亚群的聚类,不同细胞周期的细胞在各个亚群中占比不同。各个亚群的基因表达及分化能力不同,亚群5向各个方向分化的评分均较高,亚群4则均处于较低水平。结论:ScRNA-seq能够对人AF-MSC进行细致分群,精确反映各个亚群细胞间的差异。Aim:To investigate the heterogeneity of cell subgroups of cultured human amniotic fluid mesenchymal stem cells(AF-MSC)by using single cell RNA sequencing(scRNA-seq).Methods:Amniotic fluid was collected from pregnant women in the second trimester,and then isolated and cultured to obtain AF-MSC.Subsequently,AF-MSC were detected by using 10x scRNA-seq platform.The sequencing data were analyzed by filtering,comparing,quantifying,and screening.Clustering with dimensionality reduction was performed and cell cycle was determined based on gene expression level.Differential gene analysis was performed on the cell subgroups to obtain marker genes,the differentiation ability of different clusters was predicted,and the expression levels of diverse surface markers were analyzed.Results:A total of 6717 cells were detected by scRNA-seq.The cells were clustered into 7 subgroups according to gene expression levels.All the subgroups were MSC except cluster 4.The clustering of AF-MSC subgroup were affected by cell cycle,and the proportion of cells in different cell cycles was different in each cluster.The gene expression and differentiation ability of different subgroups were different,the differentiation ability of cluster 5 to all the lineages was relatively high,while cluster 4 was at a lower level.Conclusion:ScRNA-seq on AF-MSC demonstrates detailed clustering of cell subtypes and accurately reflects the differences between AF-MSC clusters.

关 键 词：单细胞RNA测序 间充质干细胞 羊水间充质细胞 异质性 

分 类 号：R318.16[医药卫生—生物医学工程]